Rapid screening method for 42 kinds of commonly used antibiotics residues in fishery water by High performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS) has been developed. Targeted antibiotics was extracted from samples on C₁₈ Solid-phase extraction cartridges. Analytical separation was performed on a C₁₈ reverse phase column by liquid chromatography with 1% acetic acid, formic acid, ammonium salt aqueous methanol solution as mobile phase. Positive ion source full scan patterns were used. The result showed that Limit of Quantity (S/N=10, LOQ) of 42 kinds of antibiotics were 0.010-0.120 μg/L. The average recovery for antibiotics was 60.7%-97.1%. The relative standard deviation (RSDs) was 0.58%-8.01%. The method is rapid and simple with high sensitivity. 60 fishery water samples were determined and sulfamethoxazole, sulfamonomethoxine, sulfapyridine, sulfachlorpyridazine, enrofloxacin were detected with the concentration from 25.6 to 106.7 ng/L. The method is sensitive, accurate and can be applied to the screening of antibiotic residues in fishery water environment.